Department of Pediatrics, Justus-Liebig-University Giessen, 35392 Giessen, Germany.
Department of Cell Biology and Cell Pathology, Philipps-University Marburg, 35032 Marburg, Germany.
Int J Mol Sci. 2023 Jan 30;24(3):2635. doi: 10.3390/ijms24032635.
Celiac disease (CD) represents a frequent autoimmune disease triggered by the ingestion of gliadin in genetically predisposed individuals. The alteration of enterocytes and brush border membrane morphology have been repetitively demonstrated, but the underlying mechanisms remain unclear. Microtubules represent a major element of the cytoskeleton and exert multiple functions depending on their tyrosination status. The aim of our study was to investigate whether posttranslational modification of microtubules was altered in the context of CD and whether this mechanism contributed to morphological changes of CD enterocytes. We examined the expression of tubulin tyrosine ligase (TTL) and vasohibin-2 (VASH2) and the level of detyrosinated and acetylated tubulin in duodenal biopsies and Caco-2 cells by immunoblot and immunofluorescence microcopy. Electron microscopy was performed to investigate the subcellular distribution of detyrosinated tubulin and brush border membrane architecture in CD biopsies and Madin-Darby Canine Kidney type II (MDCK) cells lacking TTL. CD enterocytes and Caco-2 cells stimulated with digested gliadin or IFN-y displayed a flattened cell morphology. This disturbed cellular architecture was accompanied by an increased amount of detyrosinated and acetylated tubulin and corresponding high expression of VASH2 and low expression of TTL. The altered posttranslational modification of tubulin was reversible after the introduction of the gluten-free diet. CD enterocytes and MDCK cells deficient in TTL displayed a reduced cell height along with an increased cell width and a reduced number of apical microvilli. Our results provide a functional explanation for the observed morphological alterations of the enterocytes observed in CD and provide diagnostic potential of the tyrosination status of microtubules as an early marker of villous atrophy and CD inflammation.
乳糜泻(CD)是一种由遗传易感性个体摄入麦胶蛋白引起的常见自身免疫性疾病。已经反复证明了肠细胞和刷状缘膜形态的改变,但潜在的机制仍不清楚。微管是细胞骨架的主要组成部分,根据它们的酪氨酸化状态发挥多种功能。我们的研究目的是探讨微管的翻译后修饰是否在 CD 中发生改变,以及这种机制是否有助于 CD 肠细胞的形态变化。我们通过免疫印迹和免疫荧光显微镜检查了十二指肠活检和 Caco-2 细胞中微管酪氨酸连接酶(TTL)和血管抑制素-2(VASH2)的表达以及去酪氨酸化和乙酰化微管的水平。电子显微镜用于研究 CD 活检和缺乏 TTL 的 Madin-Darby Canine Kidney 型 II(MDCK)细胞中去酪氨酸化微管和刷状缘膜结构的亚细胞分布。用消化的麦胶蛋白或 IFN-y 刺激的 CD 肠细胞和 Caco-2 细胞显示出扁平的细胞形态。这种紊乱的细胞结构伴随着去酪氨酸化和乙酰化微管的增加以及相应的 VASH2 高表达和 TTL 低表达。引入无麸质饮食后,微管的改变的翻译后修饰是可逆的。缺乏 TTL 的 CD 肠细胞和 MDCK 细胞显示出细胞高度降低,同时细胞宽度增加,顶微绒毛数量减少。我们的结果为 CD 中观察到的肠细胞形态改变提供了功能解释,并为微管的酪氨酸化状态作为绒毛萎缩和 CD 炎症的早期标志物提供了诊断潜力。